The following is a section of the Elemental Analysis Manual for Food and Related Products.

For additional information and to view other sections of the manual, visit the Elemental Analysis Manual
for Food and Related Products web page at
http://www.fda.gov/Food/FoodScienceResearch/LaboratoryMethods/ucm2006954.htm.
 Elemental Analysis Manual
for Food and Related Products

4.13 INDUCTIVELY COUPLED PLASMA-

MASS SPECTROMETRIC DETERMINATION
OF IODINE IN FOOD USING TETRAMETHYL
AMMONIUM HYDROXIDE EXTRACTION

Version 1.0 (Jan 2017)

Current Validation Status:
AOAC/ASTM: No Authors:
SINGLE LAB VALIDATION: YES Todor I.Todorov
MULTI-LAB VALIDATION: NO Patrick J. Gray

GLOSSARY

Table of Contents
4.13 INDUCTIVELY COUPLED PLASMA-MASS SPECTROMETRIC DETERMINATION OF IODINE
IN FOOD USING TETRAMETHYL AMMONIUM HYDROXIDE EXTRACTION ........................................1
4.13.1 SCOPE AND APPLICATION .........................................................................................................................2
4.13.2 SUMMARY OF METHOD ............................................................................................................................2
4.13.3 EQUIPMENT AND SUPPLIES .......................................................................................................................2
4.13.4 REAGENTS AND STANDARDS ..................................................................................................................... 4
4.13.5 EXTRACTION PROCEDURE ........................................................................................................................6
4.13.6 DETERMINATION PROCEDURE ..................................................................................................................7
4.13.7 CALCULATIONS ...................................................................................................................................... 11
4.13.8 QUALITY CONTROL ................................................................................................................................. 12
4.13.9 REPORT .................................................................................................................................................. 13
4.13.10 METHOD VALIDATION ............................................................................................................................ 13
4.13.11 REFERENCES ...................................................................................................................................... 14
FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

4.13.1 SCOPE AND APPLICATION

This method describes a procedure for the determination of iodine in foods and dietary
supplements by alkaline extraction followed by inductively coupled plasma-mass spectrometric
detection (ICP-MS).
This method should only be used by analysts familiar with the determination of trace elements
and experienced in the use of ICP-MS. The method has been validated for food, beverage and
some dietary supplement matrices.

4.13.2 SUMMARY OF METHOD

An analytical portion (0.5 to 5.0 g dependent on food composition) is mixed with tetramethyl-
ammonium hydroxide (TMAH) and a hot block extraction system at 85oC is used to extract the
available iodine. The supernatant contains extractable iodine in 1% TMAH at pH>9. The
analytical solution is analyzed using an ICP-MS. Iodine mass fraction is quantified using an
external calibration and quality controls are incorporated to ensure data quality.
Typical analytical limits were calculated per EAM §3.2 and are listed in 4.13 Table 1 but will
vary depending on the specific instrumentation, analytical portion mass, blank contamination,
sensitivity and operating conditions.

4.13 Table 1. Typical Analytical Limits

Element Symbol MBKL MBKC ASDL ASQL LOD LOQ

127
Iodine I 0.085 0.110 0.043 0.381 4.1 36.5
Based on 0.5 g analytical portion and 50 g analytical solution, 24 method blanks from 8 extractions
analyzed over 2 months (see EAM §3.2)
All analytical limits are in µg/kg

4.13.3 EQUIPMENT AND SUPPLIES

Disclaimer: The use of trade names in this method constitutes neither

endorsement nor recommendation by the U. S. Food and Drug Administration.
Equivalent performance may be achievable using apparatus and materials other
than those cited here.

(1) Inductively coupled plasma-mass spectrometer (ICP-MS)—Capable of scanning the

mass-to-charge (m/z) range 5 – 240 amu with a minimum resolution of 0.9 amu at 10%
peak height, mass flow controller for nebulizer gas.
(2) Nebulizer and spray chamber—Concentric quartz (Meinhard part number #ME2040-
54), Glass Expansion concentric (Glass expansion part number # ARG-1-QSS1) ) or
Glass Expansion MicroMist (part number # AR35-1-FM02E) and 20 mL cyclonic
spraychamber (Meinhard part number # ML151018ES, or Glass expansion part number

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

# 20-809-0261HE)
(3) Hot block extraction system—Requires uniform temperature control to at least 85 ± 2
°C. Recommended equipment is SCP science DigiPrep MS (part number #010-500-
205).
(4) Laboratory centrifuge – Capable of 3000 rpm (capable at least of max RCF of 1300 g)
and 50 mL centrifuge tubes
(5) Labware—All reusable laboratory ware must be sufficiently clean for trace metals
analysis. The recommended cleaning procedure for all laboratory ware includes
washing in special clean-rinsing laboratory detergent such as Micro-90, reagent water
rinse, soaking in 10% nitric acid and final reagent water rinse immediately before use.
Glass should not be used for dilution or storage of sample or standard solutions because
of possible contamination.
(6) Plastic labware—This includes disposable plastic laboratory ware such as autosampler
tubes and capped centrifuge tubes. Plastic bottles for solution storage should be tested
for contamination before using a particular lot with 1% TMAH rinse immediately
before use. Items can also be cleaned, dried and stored in a dust free environment for
later use. FEP, PFA, PP, LDPE or HDPE are recommended materials for bottles and
tubes. FEP, FEP coated or polypropylene spatulas should be used for sampling food
portions. Becton Dickinson polypropylene Falcon centrifuge tubes (blue cap) and Fisher
8 mL polypropylene culture tubes (Fisher part number # 14-956-7a) can be used for
sample preparation and analysis.
(7) Gloves—Use powder free vinyl or nitrile. Do not use powdered gloves or latex because
of possible contamination. Gloves manufactured for clean room use that are free from
trace metals contamination are suggested. It is good practice to put on gloves and then
rinse with reagent water to remove residual plasticizers or releasing agents before
handling clean laboratory ware or samples.
(8) Analytical balance—Capable of measuring to 0.1 mg.
(9) Top Loading balance—Capable of measuring from 0.01 g to 2500 g.
(10) Micropipettes—Air displacement micropipettes with metal free colorless disposable
plastic tips. Do not use colored tips due to possible contamination. If applicable, remove
metal tip ejector to avoid potential contamination.
(11) Clean air hood/canopy—Class 100 polypropylene metal free hoods/canopies are
recommended for sample handling.
(12) Peristaltic pump tubing—The recommended sample and internal standard peristaltic
pump tubing are orange:green (0.38 mm inner diameter). At 0.2 rev/s approximately
100 µL/min sample and 100 µL/min internal standard solutions are delivered to the
nebulizer. For higher flow nebulizers, Glass Expansion concentric, black:black (0.76
mm inner diameter) at 0.1 rev/s, delivers approximately 350 µL/min sample and 350
µL/min internal standard.
(13) Drain tubing—The recommended drain tubing is yellow:blue (1.52 mm i.d.) or larger
Fluran tubing which drains > 1000 µL/min from the spray chamber. Using less than

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

1.52 mm i.d. drain tubing will result in spray chamber flooding and instrument damage.
(14) Optional plastic syringes – general use and nonsterile, 5 or 10 mL, Luer-Loc tip.
(15) Optional PTFE syringe filter (Pall Gelman Acrodisk PTFE 1um, part number Z259926-
1PAK, or Environmental express Filtermate PTFE plunger filter).

4.13.4 REAGENTS AND STANDARDS

Reagents might be significantly contaminated with iodine. Use high purity or trace metals grade
reagents at all times. Check all new reagents for contamination prior to use. Non-quantifiable (<
ASQL) blank levels can be obtained by utilizing best laboratory practices and high purity
reagents. Blank levels < ASQL are the objective for all labs and analysts.

Safety Notes: Reagents should be regarded as potential health hazards and

exposure to these materials should be minimized. Follow universal precautions.
Wear gloves, a lab coat, and safety glasses while handling reagents.

Exercise caution when handling and dispensing concentrated strong bases. Bases
are caustic chemicals that are capable of causing severe eye and skin damage. If
acids or bases come in contact with any part of the body, quickly wash the
affected area with copious quantities of water for at least 15 minutes.

Reagents
(1) Reagent water—Water processed to meet specifications for ASTM Type-I water1.
Method validation was done using 18.2 MΩ-cm deionized water (example is Millipore
Milli-Q element system).
(2) Argon supply for instrument—High purity (99.99%) liquid argon. Argon compressed
gas tanks can also be used but is more expensive than liquid argon.
(3) High purity tetramethyl-ammonium hydroxide —25% (m/m), electronics grade
(99.9999% purity). For the method validation Alfa Aesar part number # 20932.
(4) High purity isopropanol—Electronic grade or equivalent. For the validation Fisher LC-
MS Optima grade isopropanol was used.
(5) Triton X-100 (ACS grade)

Solutions
(1) TMAH 5% (m/m)—Dilute 100 g electronic grade 25% TMAH to 500 g with reagent
water.
Recommendation: Use an empty Teflon bottle originally used for concentrated
nitric acid. To minimize contamination, dilute gravimetrically on a top loading
balance with a capacity of a least 1000 g. Tare bottle. Fill with approximately
300 mL reagent water. Note mass. Add 100 g TMAH while pouring slowly from

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

the stock bottle. Add reagent water until a total solution mass of 500 g is reached
(400 g water + 100 g TMAH). Place bottle cap on and mix.
(2) TMAH 1% (m/m)—Dilute 80 g high purity TMAH to 2,000 g with reagent water.
Recommendation: Use an empty Teflon bottle originally used for concentrated
nitric acid. To minimize contamination dilute gravimetrically on a top loading
balance with a capacity of a least 2500 g. Tare bottle. Fill with approximately
1000 mL reagent water. Note mass. Add 80 g TMAH while pouring slowly from
the stock bottle. Add reagent water until a total solution mass of 2000 g is
reached (1920 g water + 80 g TMAH). Place bottle cap on and mix.
(3) Internal standard solution (ISTD)—Multi-element solution prepared by diluting an
appropriate mass of stock standard. ISTD matrix is 1% TMAH, 6% isopropanol, 0.01%
triton X-100. The presence of isopropanol will help equalize iodine sensitivity due to
residual carbon remaining in solution after the extraction. The dilution factor of the
internal standard solution is 1:1 if the autosampler and internal standard peristaltic
pump tubes are equal inner diameter. The analytical solution pumped into the nebulizer
will be approximately 3% isopropanol.
a. The exact mass fraction is not as important as maintaining the same mass
fraction over an analytical run. Since the element mass fraction is only
approximate, the solution may be prepared volumetrically.
b. ISTD elements and suggested mass fractions: 2±0.5 µg/kg Rh, 20±2 µg/kg Te.
c. ISTD solution must be prepared daily because of instability of Rh in alkaline
solutions longer than 48 h.
(4) Suggested Tuning Solution — 1 µg/kg iodine solution in 1% TMAH used to tune and
optimize instrument. Typical sensitivity for 1 µg/kg I should be better than 50,000
cps/ppb.
The method specifies sample tubing and internal standard tubing to be equal
diameter, diluting tune solution by half.

Calibration Standard Solutions

(1) Analyte stock standard solutions — commercially prepared single element NIST
traceable standard solutions prepared specifically for plasma mass spectrometric or ion
chromatography analysis should be used. Due to the low mass fractions of solutions
required for ICP-MS standardization, starting from 10 mg/kg stock solutions is
recommended to minimize the number of dilutions and intermediate solutions.
a. Standards can be purchased on a mass/mass basis to eliminate density correction
factors. If standards are mass/volume, a density correction will be necessary
(refer to EAM §3.4.4 for gravimetric standard solution preparation).
b. Stock standard solutions must be used prior to manufacturer’s expiration date
(typically 12 – 18 months from time of purchase). Be aware that stock solutions
may slowly become more concentrated over time due to transpiration of water

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

vapor through the bottle material and loss while the bottle is uncapped.
(2) Standard solutions—Dilute stock standard with 1 % TMAH to prepare iodine standards.
Depending on the mass fraction of the stock standard the use of serial dilutions is
recommended in preparing the calibration standards. Store in Teflon® FEP, PP or HDPE
bottles. A minimum of 4 calibration standard levels should be used for calibration (4.13
Table 2).

4.13 Table 2. Example of calibration curve standard mass fractions

Level 0 Level 1 Level 2 Level 3 Level 4 Level 5 Level 6 Level 7
Analyte
(µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg) (µg/kg)
I 0 0.010 0.050 0.200 1.0 5.0 20.0 50.0

(3) Standard blank— 1 % TMAH.

(4) Initial (independent) calibration verification (ICV) — Dilute an appropriate volume of
stock iodine solution obtained from a different (second) source gravimetrically with 1 %
TMAH so that the analyte mass fraction will be at the approximate midpoint of the
calibration curve.
(5) Continuing calibration verification (CCV) — Use a mid-level standard.

4.13.5 EXTRACTION PROCEDURE

Terms and definitions:
(1) An “extraction batch” is defined as digests from the vessels in a single tray in the same
extraction program at the same time. The SCP Science DigiPrep extraction batch will
have up to 48 vessels.
(2) An “analytical run” is defined as the total number of analytical solutions analyzed
during a single sequence following tuning and optimization and with one calibration.
An analytical run may contain analytical solutions from more than one extraction batch.
The following operations should be performed in a clean environment to reduce contamination.
An exhausting hood must be used when working with TMAH.
Typically, only the edible portions of foods are analyzed. However, if an assignment requires
mass fractions as a function of dry mass, dry a minimum of 10 g of the homogenized, ground
samples in a laboratory oven at 85 °C until a constant mass is obtained. Standard reference
materials (SRM) should be dried according to the manufacturer’s recommendations. Calculate
the moisture content of the original sample. Store dried samples in a desiccator.
Food preparation and homogenization procedures are found in EAM §2.1 through EAM §2.2.2.

Extraction Procedure using DigiPrep hot block

(1) Weigh each 50 mL centrifuge tube and record mass with cap.

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

(2) In each extraction batch, a minimum of two method blanks must be included to check
contamination from the vessels. The method blanks should be placed in random vessels.
(3) Place 0.5 g analytical portion into clean centrifuge tube and record mass of tube and
sample.
a. Less than the maximum mass should be used for samples high in salt content.
b. For most beverage and liquid samples, use an analytical portion mass of 5 g.
c. Use 0.5 g reagent water for method blanks (MBK) and optional fortified method
blanks (FMB).
d. For dry samples and dry SRM materials adding 1 g of reagent water can help
control reactions during the extraction.
(4) Pipette 10 mL of 5% TMAH into centrifuge tube, washing down any material on walls.
Using a bottle top dispenser is suggested.
(5) Vortex each centrifuge tube containing sample and TMAH for 1 min.
(6) Place capped samples on hot block. The hot block extraction of iodine temperature
program contains a 30 min ramp to 85 °C and 150 min hold at 85 °C.
(7) After vessels have cooled to less than 30 °C (approximately 2 h if the samples are left to
cool on the hot block) remove tubes and vortex for 1 minute.
(8) Add reagent water to 50 mL mark, vortex for 1 minute, weight samples and record final
mass.
(9) Centrifuge samples at 3000 rpm for 3 min. Collect supernatant and analyze by ICP-MS
using the procedure listed below.
(10) (optional step) If large amount of solids are present, filtering of the supernatant can
prevent clogging of the nebulizer or sample probe. Uptake 5 mL of supernatant and
filter into a clean sample tube for analysis. Discard 1 mL of the supernatant to prevent
contamination from the filter.

4.13.6 DETERMINATION PROCEDURE

The method was developed using Agilent 7700x and Thermo Element 2 mass spectrometers. As
there are no significant interferences for iodine at m/z 127, the Agilent is used in “no cell gas” or
vented mode and the Thermo Element 2 is used in low resolution mode. Internal standards are
used to help compensate for matrix effects and general instrumental drift. References to
sensitivity, cps and % RSD refer to the Agilent 7700x instrument.

Instrument Setup
(1) See EAM §3.6.1.4 for additional details on ICP-MS.
Use a separate sample introduction system (sample probe, peri-pump tubing, nebulizer,
spray chamber, injector, torch and cones) for alkaline solutions (TMAH). The sample
introduction system need to be rinsed with 1% TMAH or 1% optima grade NH4OH for
a minimum of 4 hours before the first use for iodine measurements in order to wash out

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

iodine from all glass and plastic components. Once the ICP-MS is switched to alkaline
sample introduction it is recommended that several batches are run before switching to
acid solution mode. Frequent acid to base switching may require a prolonged iodine
washout in order to obtain the above mentioned iodine count rates.
(2) Perform manufacturer recommended instrument start up procedure or laboratory
specific procedures.
May include the following checks: Ar supply pressure, backing pump oil
condition, sufficient exhaust flow, and peristaltic pump tubing condition.
(3) Ignite plasma and perform initiation procedures as instructed in the owner's manual.
a. Fill a rinse bottle with 1% TMAH. Send autosampler probe to the rinse bottle
while instrument is warming up. Place internal standard line in tube containing
reagent water during warm up. Rinse and warmup the instrument for a minimum
of 1 hour.
b. Program autosampler sequence table to run standards and samples of the batch.
(4) Set up method to include analytes and internal standard elements as shown in 4.13
Table 3.

4.13 Table 3. ICP-MS Instrument Parameters

Recommended Minimum
Monitored Internal isotope for Analysis
Element Integration
Isotope Standard reporting Mode
Time (sec)
127 103 127
Iodine I Rh or 125Te I 0.3 No gas
103
Rhodium Rh — 0.3 No gas
125
Tellurium Te — 0.3 No gas

a. Use 3 points per peak and at least 3 replicates for integration. Use the mean of
the integrations for reporting.
b. Be sure there is adequate rinse time programmed in between samples. Program
the autosampler probe to go to the rinse station for at least 10 seconds after
analyzing an analytical solution and then to the rinse bottle filled with 1%
TMAH. The rinse time must be great enough so that a standard blank solution
produces stable iodine baseline signal. A minimum of a three minute rinse is
recommended.
c. An “intelligent rinse” or “smart rinse” feature may be used if so equipped.
Analyte levels must return to within 10% RPD of the calibration blank cps levels
before moving onto the next analytical solution.
(5) Optimize instrument
a. Configure the tune to monitor 103Rh, 125Te and 127I. Introduce calibration blank
solution. Pump speed during tuning and analyses should be set at 0.1 rev/s.
Typical sensitivity to be achieved in an Agilent 7700x system is 300,000 cps for

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

103
Rh, and 100,000 cps for 125Te. The background for iodine in a blanks solution
should be at below 6,000 cps.

Note: During tuning, the internal standard tubing is placed in the ISTD solution
containing 1 % TMAH, 0.01% triton-x100 and 6% IPA.

b. Tune for highest stability while maintaining optimal sensitivity for the m/z 125
and 103, and lowest cps at m/z 127.
c. Save updated and optimized tune file
d. Check instrument performance using a 1 mg/kg I standard. Typical sensitivity to
be achieved at 0.1 rev/s, black/back tubing is 50,000 cps with 1-2% RSD.
e. Precision Check: Demonstrate instrument stability by analyzing a midrange
iodine standard solution (e.g. CCV). The resulting relative standard deviation
(RSD) of ion signals must be ≤10%. If RSD > 10%, determine and correct
problem before standardization. Stability problems are usually related to sample
introduction.

Determination of Analyte Mass Fraction Using External Standard Calibration Curve

An example of a typical analytical sequence is shown in 4.13 Table 4
Calibrate using the standard blank and at least four iodine standards. A calibration blank is used
as a point on the calibration curve (0 µg/kg calibrant). Additionally a high standard check at or
around 50 µg/kg iodine should be analyzed as a sample to ensure linearity to 50 µg/kg iodine.
The high standard linearity check should be within 10 % of the calculated mass fraction.
(1) Use linear regression with blank offset and no weighing factor.
(2) Check standardization performance
a. Linear regression correlation coefficient (r) (intensity - (analyte
counts/sec):(internal standard counts/sec)) versus mass fraction) is ≥ 0.9975.
b. Analyze initial calibration verification (ICV) to verify standardization. Recovery
must be 100 ± 10%.
c. Analyze a high standard check at or around 50 µg/kg iodine as a sample to
ensure linearity to 50 µg/kg iodine. The high standard linearity check should be
within 10 % of the calculated mass fraction.
(3) Check instrument measurement performance and analyze analytical solutions
a. Interpolate analyte mass fraction from standard curve. Start samples analysis
sequence and analyze the highest standard, standard blank and ICV in that order.
This will verify proper autosampler rinse time and valid calibration curve.
b. Continuing calibration verification (CCV) must be analyzed at every 10 samples
and at the end of the analytical run. Recovery must be 100 ± 10%.
c. RSD of the measurements of replicate integrations must be ≤ 10% for all

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

solutions when instrument response > ASQL.

d. Continuing calibration blank (CCB) analyzed at a frequency of 10% and at the
end of the analytical run. CCB solutions should be ≤ ASQL.
e. Analytical solutions producing mass fractions which are greater than the high
linearity check solution should be diluted with 1% TMAH and re-analyzed at a
level falling within the lowest non-zero standard and the high standard.
(4) Suppression or enhancement of internal standard isotope response may indicate a matrix
effect is present. Monitor internal standard signals and dilute any analytical solution

4.13 Table 4. Example of Typical Analytical Sequence

Grouping Solution QC Criteria
tune report RSD, sensitivity
precision check (n > 10) ≤10% RSD
calibration standards r ≥ 0.9975
standard blank ≤ASQL
Calibration high standard solution 90% - 110% of calculated mass fraction
standard blank ≤ASQL (memory check)
ICV 90% - 110% recovery
MBK 1 ≤MBKC
2/3 of MBKs
MBK 2 ≤MBKC
≤MBKC
MBK 3 ≤MBKC
SRM 80% - 120% recovery
sample 1 ≤10% instrument RSD, < high cal. std
sample 1 duplicate
sample 1 FAP 80% - 120% recovery
sample 2
Unknowns - sample 3
Set 1 sample 4
sample 5
sample 6
sample 7
CCV 90% - 110%
CCB ≤ ASQL
sample 8 ≤ 10% instrument RSD, < high cal. std
sample 8 duplicate
sample 8 FAP
sample 9
sample 10
Unknowns -
Set 2 sample 11
sample 12
sample 13
sample 14
CCV 90% - 110%
CCB ≤ ASQL
Precision required: All solutions must be ≤10% RSD when analyte ≥ASQL.

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

(5) where the internal standard signal differs by more than 40% from the standard blank.
Use 1 % TMAH for diluent. Rh is suggested to be used as a primary internal standard
element.
(6) Elevated internal standard isotope response may indicate the presence of the internal
standard element in the sample or an interference on the internal standard isotope. If the
internal standard signal is greater than 140% of the standard blank, choose a different
internal standard and reprocess the data.
(7) Analyze duplicate analytical portions every 10 samples. The duplicate analytical
portions must have relative percent difference < 20% when analyte mass fractions are >
LOQ. If it fails, repeat analysis of the duplicate portion. If it fails again, re-digest and
re-analyze. Analyze duplicate analytical portions for each sample type in a run (if all
samples have similar compositions, only one duplicate portion must be analyzed for
every 10 samples). It is highly recommended that duplicate analytical portions be
analyzed for every food sample if feasible.
(8) At least one fortified analytical portion (FAP) should be included in each analytical run
and if more than 10 samples are extracted, an FAP should be included for every 10
samples. Fortification recoveries are described in EAM §3.4.1. The marginal method of
calculating percent recovery is used for fortification recovery calculations2.
a. FAP preparation: Spike 50-300% of the native elemental mass fraction,
FAP % marginal recovery: 80 - 120%. If it fails, re-analyze one time. If
the FAP fails again, re-digest and re-analyze.

4.13.7 CALCULATIONS
Calculate the mass fraction of the analyte in the analytical portion according to the formula
𝜇𝜇𝑔𝑔 𝑀𝑀
𝑀𝑀𝑎𝑎𝑎𝑎𝑎𝑎 𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓 � � = [(𝑆𝑆 × 𝐷𝐷𝐷𝐷) − 𝑀𝑀𝑀𝑀𝑀𝑀𝐿𝐿 ] ×
𝑘𝑘𝑔𝑔 𝑚𝑚 × 𝑀𝑀𝑀𝑀𝐷𝐷
where
S = mass fraction of analyte in analytical solution (or diluted analytical
solution) (µg/kg)
MBKL = laboratory MBK (mg/kg) (subtract if MBK is greater than ASQL) (EAM
§3.6.2)
DF = dilution factor (1 if analytical solution not diluted) (EAM §3.4.3)
MCF = mass correction factor (1 if no water or other solvent was added to aid
homogenization) (EAM §3.4.6)
M = Mass (g) of analytical solution (usually 50 – 100 g)
m = mass of analytical portion (g)

Round calculated mass fraction to at most 3 significant figures. Mass fractions may be converted
to other convenient units (e.g., µg/kg, ng/g for solids or ng/L for liquids).
Calculate the marginal recovery (%) in the fortified analytical portion according to the formula

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

𝐶𝐶
% 𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅𝑅 =

where
Cx+s = concentration determined in spiked sample (µg/kg)
Cx = concentration determined in unspiked sample (µg/kg)
Cs = concentration of spiking solution (µg/kg)
Ms = mass of spiking solution added to analytical portion (g)
Mx = mass of analytical portion (g)

4.13.8 QUALITY CONTROL

The following is the minimum number of quality control samples analyzed with each analytical
run:
• 1 certified reference material (SRM/CRM)
Match reference material matrix to the food matrix. In-house RMs are acceptable only if
no traceable RM is available and the in-house RM is well characterized.
• 1 fortified analytical portion (FAP) per sample type per every 10 samples
• 2 method blanks (MBKs)
• 1 duplicate sample preparation for every 10 samples
• Replicate analytical portions should be analyzed for each sample whenever sample non-
homogeneity may be an issue.

Reference Material
Control limits for True Value recovery of reference materials are 100 ± 20% or within mass
fraction uncertainty (converted to percent relative uncertainty) supplied on certificate, whichever
is greater. Traceable standard or certified reference materials (CRM/SRM) should be used when
available, for example NIST 1549a (non-fat milk), NIST 1566a (oyster tissue).

FAP Recovery
Control limit for FAP marginal recovery is 80 – 120%.

Method Blanks (MBK)

Minimum of 2 MBKs analyzed in an analytical run and mass fraction of both MBKs must be ≤
MBKC. If 3 or more MBKs are analyzed then at least two-thirds of MBKs must be ≤ MBKC.

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

Relative Percent Difference (RPD) of Replicate Analytical Portions

Control limit for RPD is 20% for analyte levels > LOQ. (EAM §3.4.5)

4.13.9 REPORT
Report results only when all the quality control criteria for a batch have been satisfactorily met.
Report results that are ≥LOQ as the mass fraction determined followed by the units of
measurement. Report results that are ≥LOD and <LOQ as the mass fraction determined followed
by the units of measurement and the “Trace” data qualifier that indicates analyte is present at a
trace level that is below the limit of reliable quantification. Trace values are documented by a
“TR” after the result. Report results that are <LOD as 0 followed by the units of measurement
and the qualifier that indicates analyte is below the level of reliable detection or is not detected
(ND).

Example: LOQ = 10 µg/kg; LOD =3 µg/kg. Levels found for three different
samples were 11 µg/kg, 5 µg/kg and 2 µg/kg.

11 µg/kg is ≥LOQ; report 11 µg/kg

5 µg/kg is ≥LOD but also <LOQ; report 5 µg/kg (TR)

2 µg/kg is <LOD; report 0 µg/kg (ND)

If an analytical portion is analyzed in duplicate and one replicate mass fraction is >LOD but
<LOQ and the other replicate is >LOQ, average the two results and report the measurements
using the rules shown above.

4.13.10 METHOD VALIDATION

Single lab validation
EAM 4.13 was validated following the guidelines set forth in Guidelines for the Validation of
Chemical Methods for the FDA Foods Program3 and exceeds the standard method performance
requirements approved by the AOAC Stakeholder Panel on Strategic Food Analytical Methods4.
The method was validated by analyses of reference materials and fortified analytical portions for
accuracy and replicate portions for precision. Reference materials used for validation are listed in
single laboratory validation publication5.
Analyses were performed on 21 different foods that were similar to those collected in FDA’s
Total Diet Study purchased from local grocers. Foods were prepared as described and analyzed
several (N ≥ 3) times. Fortified analytical portions (3 spike levels each) were also prepared and
analyzed. Method validation results are shown in Todorov et al 20165.
Uncertainty.
A detailed discussion of method uncertainty is presented in EAM EAM §3.3. This method
conforms to the information contained in that discussion. Derivation of an estimated uncertainty
specific to an analysis is discussed EAM §3.3.2.

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FDA Elemental Analysis Manual (Section 4.13 Iodine ICP-MS method)

Multi-lab validation.
[Under development]

4.13.11 REFERENCES
(1) ASTM International (2006) ASTM D 1193-06, Standard Specification for Reagent Water. Available
from ASTM (http://www.astm.org).
(2) Official Methods of Analysis of AOAC INTERNATIONAL (2005) 18th Ed., AOAC International,
Gaithersburg MD, USA, Appendix D: Guidelines for Collaborative Procedures To Validate
Characteristics of a Method of Analysis.
(3) U.S. Food and Drug Administration (2015) Guidelines for the Validation of Chemical Methods for the
FDA Foods Program, Version 2. Available from FDA
(http://www.fda.gov/downloads/ScienceResearch/FieldScience/UCM273418.pdf).
(4) AOAC International (2013) AOAC SMPR 2012.007 – Standard Method Performance Requirements for
Heavy Metals in a Variety of Foods and Beverages, J. AOAC Int. 96, 704.
(5) Todorov TI, Gray PG, 2016, Analysis of iodine in food samples by inductively coupled plasma-mass
spectrometry, Food Additives and Contaminants, Part A, 33, 282

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